Novel system for detecting SARS coronavirus nucleocapsid protein using an ssDNA aptamer.
Identifieur interne : 001473 ( PubMed/Corpus ); précédent : 001472; suivant : 001474Novel system for detecting SARS coronavirus nucleocapsid protein using an ssDNA aptamer.
Auteurs : Seong-Je Cho ; Hye-Min Woo ; Ki-Sun Kim ; Jong-Won Oh ; Yong-Joo JeongSource :
- Journal of bioscience and bioengineering [ 1347-4421 ] ; 2011.
English descriptors
- KwdEn :
- Aptamers, Nucleotide (chemistry), Aptamers, Nucleotide (metabolism), Base Sequence, Blotting, Western, DNA, Single-Stranded (chemistry), DNA, Single-Stranded (metabolism), Enzyme-Linked Immunosorbent Assay, Humans, Molecular Sequence Data, Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (isolation & purification), SARS Virus (genetics), SARS Virus (isolation & purification), SELEX Aptamer Technique (methods), Sensitivity and Specificity, Sequence Alignment, Severe Acute Respiratory Syndrome (diagnosis).
- MESH :
- chemical , chemistry : Aptamers, Nucleotide, DNA, Single-Stranded.
- chemical , genetics : Nucleocapsid Proteins.
- chemical , isolation & purification : Nucleocapsid Proteins.
- chemical , metabolism : Aptamers, Nucleotide, DNA, Single-Stranded.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- methods : SELEX Aptamer Technique.
- Base Sequence, Blotting, Western, Enzyme-Linked Immunosorbent Assay, Humans, Molecular Sequence Data, Sensitivity and Specificity, Sequence Alignment.
Abstract
The outbreak of severe acute respiratory syndrome (SARS) in 2002 affected thousands of people and an efficient diagnostic system is needed for accurate detection of SARS coronavirus (SARS CoV) to prevent or limit future outbreaks. Of the several SARS CoV structural proteins, the nucleocapsid protein has been shown to be a good diagnostic marker. In this study, an ssDNA aptamer that specifically binds to SARS CoV nucleocapsid protein was isolated from a DNA library containing 45-nuceotide random sequences in the middle of an 88mer single-stranded DNA. After twelve cycles of systematic evolution of ligands by exponential enrichment (SELEX) procedure, 15 ssDNA aptamers were identified. Enzyme-linked immunosorbent assay (ELISA) analysis was then used to identify the aptamer with the highest binding affinity to the SARS CoV nucleocapsid protein. Using this approach, an ssDNA aptamer that binds to the nucleocapsid protein with a K(d) of 4.93±0.30nM was identified. Western blot analysis further demonstrated that this ssDNA aptamer could be used to efficiently detect the SARS CoV nucleocapsid protein when compared with a nucleocapsid antibody. Therefore, we believe that the selected ssDNA aptamer may be a good alternative detection probe for the rapid and sensitive detection of SARS.
DOI: 10.1016/j.jbiosc.2011.08.014
PubMed: 21920814
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pubmed:21920814Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Novel system for detecting SARS coronavirus nucleocapsid protein using an ssDNA aptamer.</title><author><name sortKey="Cho, Seong Je" sort="Cho, Seong Je" uniqKey="Cho S" first="Seong-Je" last="Cho">Seong-Je Cho</name><affiliation><nlm:affiliation>Department of Bio and Nanochemistry, Kookmin University, Seoul 136-702, Republic of Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Woo, Hye Min" sort="Woo, Hye Min" uniqKey="Woo H" first="Hye-Min" last="Woo">Hye-Min Woo</name></author><author><name sortKey="Kim, Ki Sun" sort="Kim, Ki Sun" uniqKey="Kim K" first="Ki-Sun" last="Kim">Ki-Sun Kim</name></author><author><name sortKey="Oh, Jong Won" sort="Oh, Jong Won" uniqKey="Oh J" first="Jong-Won" last="Oh">Jong-Won Oh</name></author><author><name sortKey="Jeong, Yong Joo" sort="Jeong, Yong Joo" uniqKey="Jeong Y" first="Yong-Joo" last="Jeong">Yong-Joo Jeong</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2011">2011</date><idno type="RBID">pubmed:21920814</idno><idno type="pmid">21920814</idno><idno type="doi">10.1016/j.jbiosc.2011.08.014</idno><idno type="wicri:Area/PubMed/Corpus">001473</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001473</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Novel system for detecting SARS coronavirus nucleocapsid protein using an ssDNA aptamer.</title><author><name sortKey="Cho, Seong Je" sort="Cho, Seong Je" uniqKey="Cho S" first="Seong-Je" last="Cho">Seong-Je Cho</name><affiliation><nlm:affiliation>Department of Bio and Nanochemistry, Kookmin University, Seoul 136-702, Republic of Korea.</nlm:affiliation></affiliation></author><author><name sortKey="Woo, Hye Min" sort="Woo, Hye Min" uniqKey="Woo H" first="Hye-Min" last="Woo">Hye-Min Woo</name></author><author><name sortKey="Kim, Ki Sun" sort="Kim, Ki Sun" uniqKey="Kim K" first="Ki-Sun" last="Kim">Ki-Sun Kim</name></author><author><name sortKey="Oh, Jong Won" sort="Oh, Jong Won" uniqKey="Oh J" first="Jong-Won" last="Oh">Jong-Won Oh</name></author><author><name sortKey="Jeong, Yong Joo" sort="Jeong, Yong Joo" uniqKey="Jeong Y" first="Yong-Joo" last="Jeong">Yong-Joo Jeong</name></author></analytic><series><title level="j">Journal of bioscience and bioengineering</title><idno type="eISSN">1347-4421</idno><imprint><date when="2011" type="published">2011</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Aptamers, Nucleotide (chemistry)</term><term>Aptamers, Nucleotide (metabolism)</term><term>Base Sequence</term><term>Blotting, Western</term><term>DNA, Single-Stranded (chemistry)</term><term>DNA, Single-Stranded (metabolism)</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Humans</term><term>Molecular Sequence Data</term><term>Nucleocapsid Proteins (genetics)</term><term>Nucleocapsid Proteins (isolation & purification)</term><term>SARS Virus (genetics)</term><term>SARS Virus (isolation & purification)</term><term>SELEX Aptamer Technique (methods)</term><term>Sensitivity and Specificity</term><term>Sequence Alignment</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Aptamers, Nucleotide</term><term>DNA, Single-Stranded</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Aptamers, Nucleotide</term><term>DNA, Single-Stranded</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>SELEX Aptamer Technique</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term><term>Blotting, Western</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Humans</term><term>Molecular Sequence Data</term><term>Sensitivity and Specificity</term><term>Sequence Alignment</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The outbreak of severe acute respiratory syndrome (SARS) in 2002 affected thousands of people and an efficient diagnostic system is needed for accurate detection of SARS coronavirus (SARS CoV) to prevent or limit future outbreaks. Of the several SARS CoV structural proteins, the nucleocapsid protein has been shown to be a good diagnostic marker. In this study, an ssDNA aptamer that specifically binds to SARS CoV nucleocapsid protein was isolated from a DNA library containing 45-nuceotide random sequences in the middle of an 88mer single-stranded DNA. After twelve cycles of systematic evolution of ligands by exponential enrichment (SELEX) procedure, 15 ssDNA aptamers were identified. Enzyme-linked immunosorbent assay (ELISA) analysis was then used to identify the aptamer with the highest binding affinity to the SARS CoV nucleocapsid protein. Using this approach, an ssDNA aptamer that binds to the nucleocapsid protein with a K(d) of 4.93±0.30nM was identified. Western blot analysis further demonstrated that this ssDNA aptamer could be used to efficiently detect the SARS CoV nucleocapsid protein when compared with a nucleocapsid antibody. Therefore, we believe that the selected ssDNA aptamer may be a good alternative detection probe for the rapid and sensitive detection of SARS.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">21920814</PMID><DateCompleted><Year>2012</Year><Month>04</Month><Day>23</Day></DateCompleted><DateRevised><Year>2020</Year><Month>04</Month><Day>02</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1347-4421</ISSN><JournalIssue CitedMedium="Internet"><Volume>112</Volume><Issue>6</Issue><PubDate><Year>2011</Year><Month>Dec</Month></PubDate></JournalIssue><Title>Journal of bioscience and bioengineering</Title><ISOAbbreviation>J. Biosci. Bioeng.</ISOAbbreviation></Journal><ArticleTitle>Novel system for detecting SARS coronavirus nucleocapsid protein using an ssDNA aptamer.</ArticleTitle><Pagination><MedlinePgn>535-40</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.jbiosc.2011.08.014</ELocationID><Abstract><AbstractText>The outbreak of severe acute respiratory syndrome (SARS) in 2002 affected thousands of people and an efficient diagnostic system is needed for accurate detection of SARS coronavirus (SARS CoV) to prevent or limit future outbreaks. Of the several SARS CoV structural proteins, the nucleocapsid protein has been shown to be a good diagnostic marker. In this study, an ssDNA aptamer that specifically binds to SARS CoV nucleocapsid protein was isolated from a DNA library containing 45-nuceotide random sequences in the middle of an 88mer single-stranded DNA. After twelve cycles of systematic evolution of ligands by exponential enrichment (SELEX) procedure, 15 ssDNA aptamers were identified. Enzyme-linked immunosorbent assay (ELISA) analysis was then used to identify the aptamer with the highest binding affinity to the SARS CoV nucleocapsid protein. Using this approach, an ssDNA aptamer that binds to the nucleocapsid protein with a K(d) of 4.93±0.30nM was identified. Western blot analysis further demonstrated that this ssDNA aptamer could be used to efficiently detect the SARS CoV nucleocapsid protein when compared with a nucleocapsid antibody. Therefore, we believe that the selected ssDNA aptamer may be a good alternative detection probe for the rapid and sensitive detection of SARS.</AbstractText><CopyrightInformation>Copyright © 2011 Elsevier B.V. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Cho</LastName><ForeName>Seong-Je</ForeName><Initials>SJ</Initials><AffiliationInfo><Affiliation>Department of Bio and Nanochemistry, Kookmin University, Seoul 136-702, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Woo</LastName><ForeName>Hye-Min</ForeName><Initials>HM</Initials></Author><Author ValidYN="Y"><LastName>Kim</LastName><ForeName>Ki-Sun</ForeName><Initials>KS</Initials></Author><Author ValidYN="Y"><LastName>Oh</LastName><ForeName>Jong-Won</ForeName><Initials>JW</Initials></Author><Author ValidYN="Y"><LastName>Jeong</LastName><ForeName>Yong-Joo</ForeName><Initials>YJ</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D023362">Evaluation Study</PublicationType><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. 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MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D001483" MajorTopicYN="N">Base Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015153" MajorTopicYN="N">Blotting, Western</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004277" MajorTopicYN="N">DNA, Single-Stranded</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004797" MajorTopicYN="N">Enzyme-Linked Immunosorbent Assay</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008969" MajorTopicYN="N">Molecular Sequence Data</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D019590" MajorTopicYN="N">Nucleocapsid Proteins</DescriptorName><QualifierName UI="Q000235" 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